• Title/Summary/Keyword: cytochemical analysis

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Effects of Wax Gourd Extracts on Adipocyte Differentiation and Uncoupling Protein Genes(Ucps) Expression in 3T3-Ll Preadipocytes

  • Kang, Keun-Jee;Kwon, So-Young
    • Nutritional Sciences
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    • v.6 no.3
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    • pp.148-154
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    • 2003
  • Although various raw plant materials have been demonstrated to exert anti-obesity effects to a greater or lesser extent in both humans and animals when they are used to supplement the diet, it has not been shown extensively that they influence adipocyte cell differentiation involving lipid metabolic gene expressions. Using a well-established 3T3-L1 preadipocyte differentiation system, we decided to look into molecular and cellular event occurring during adipocyte differentiation when raw plant materials aye included in the process, in an effort to demonstrate the potential use of a screening system to define the functions of traditionally well-known materials. To these ends, the effects of ethanol (EtOH) or EtOH/distilled water (DW) extracts of Wax Gourd were examined using cytochemical and molecular analyses to determine whether components of the extracts modulate adipocyte differentiation of 3T3-Ll preadipocytes in vitro. The cytochemical results demonstrated that EtOH or EtOH/DW extracts did not affect lipid accumulation and cell proliferation, although the degree of lipid accumulation was influenced slightly depending on the extract. EtOH extract was highly effective in apoptotic induction during differentiation of 3T3-Ll preadipocytes (p<0.05). Reverse transcription-polymerase chain reaction (RT-PCR) analysis of lipoprotein lipase (LPL), Uncoupling protein (Ucp) 2, 3 and 4 also showed that while LPL expression was not influenced, Ucp2, 3 and 4 were up regulated in the EtOH extract-treated group and down regulated in the EtOH/DW extract-treated group. These changes in gene expressions suggest that the components in different fractions of Wax Gourd extracts may modulate lipid metabolism by either direct or indirect action. Taking these results together, it was concluded that molecular and cellular analyses of adipocyte differentiation involving lipid metabolic genes should facilitate understanding of cellular events occurring during adipocyte differentiation. Furthermore, the experimental scheme and analytical methods used in this study should provide a screening system for the functional study of raw plant materials in obesity research.

Cytochemical Observation of Volutin Granules and Activities of Tripolyphosphatase and Polyphosphatase in Saccharomyces uvarum (효모 세포의 Tripolyphosphatase와 Polyphosphatase 활성도 및 Volutin 과립의 세포학적 관찰)

  • Lee, Ki-Sung;Choi, Yong-Keel
    • The Korean Journal of Mycology
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    • v.13 no.3
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    • pp.141-148
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    • 1985
  • To investigate cellular regulation of phosphate metabolism between catabolically repressed and derepressed states in Saccharomyces uvarum, the activities of polyphosphatases, the analysis of polyphosphate and cytochemical observation of volutin granules were examined according to the culture phase and under various phosphate concentrations. As the results, tripolyphosphatase activity was increased more than six-fold during catabolic repression as compared with those of catabolic derepression and the polyphosphatase activity increased at the time of maximal accumulation of acid insoluble polyphosphate 'B'. Of the low molecular weight polyphosphates, tripolyphosphate was mainly detected by thin layer chromatography. When the synthesis of volutin granules in derepressed cells was observed cytochemically, acid insoluble polyphosphate localizing at the cell wall was primarily synthesized and then transferred into the cytoplasm, nucleus and/or vacuole.

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Degenerative Changes of Myocytes Induced by Restriction of Flight in Doves (비행운동 제한에 따른 비둘기 심근세포의 퇴행성 변화)

  • 문혜정;이용덕;박원학
    • Biomedical Science Letters
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    • v.6 no.4
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    • pp.269-279
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    • 2000
  • This present study was investigated to elucidate degenerative changes according to the change of habitual environment on the myocytes of doves by restricting them from flight that is instinct behavior of this animal and strong exercise. To restrict doves from flight, they were confined in the cage (1 $m^3$) for 2 months. After this period, the myocardium of the experimental group was compared to that of wild doves in the ultrastructural and cytochemical ways. In addition, stereological changes were also examined. The results were as followings: 1. The body weight of the confined experimental groups was higher than that of the wild doves, but the ratios of the pectoral muscle/body weight (p<0.05) and the heart/body weight were lower. 2. At the ultrastructural level, the myocardium of confined doves appeared as wavy fibers in the smaller area than in the myocardium of wild doves. Also, the length of sarcomeres was longer in the confined doves. The number of sarcoplasmic reticulum and capillary was smaller in the myocardium of confined doves. 3. Cytochemical examinations showed that the activities of cytochrome oxidase were lowered in the confined doves. 4. Stereological analysis revealed that the density of myofibrils was greater in the confined doves. In contrast the volume density of sarcoplasmic reticulum (p<0.05) and the surface density of mitochondrial inner membrane (p<0.05) was lower in the confined doves, while the numerical density of mitochondrial inner membrane was higher (p<0.05). These results suggest that even the short period of restricted exercise can induce negative effects on the functions of myocytes of doves that are adapted for the strong exercise such as flight. Therefore, the maintenance of prolonged exercise seems to be one of the important factors that are critical to retain the functions of myocardium.

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Ultrastructure of Cell Wall in the Suppressor Mutant of Null Pigmentation (SU-NPG) of Aspergillus nidulans (Aspergillus nidulans 색소결핍 억제돌연변이주의 세포벽 미세구조)

  • 정윤신
    • Journal of Life Science
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    • v.14 no.1
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    • pp.45-50
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    • 2004
  • To investigate the effect of the cell wall on the pigmentation and branching in Aspergillus nidulans, ultrastructure of cell wall in suppressor mutant of the null pigmentation (SU-NPG, SU602) has been examined. Scanning electron microgrphs (SEM) revealed that the most outer layer of conidia wall peeled off in SU-NPG on day 6 from the complete conidiation. They also showed that hyphal growth and branching were not well developed in SU-NPG. Transmission electron micrographs (TEM) showed that the plasma membrane was not crenulated and the hyphal wall was thick in SU-NPG. These results indicated that the ultrastructure of cell wall in SU-NPC might be modified. Cytochemical analysis showed that the cell wall in SU-NPG was differentiated into Cl, C3, C2 and C4 layer in conidia and H1, H3, H2 and H4 layer in hyphae. C3 layer and H3 layer existed in SU-NPG. The increment of the diameter in SU-NPG hyphae might be caused by the thickness of H3 layer. These results suggest that SU-NPG may have an immature but the differentiated structure for the pigmentation in cell wall.

Isolated Spinal Neurocysticercosis : Unusual Ocular Presentation Mimicking Pseudotumor Cerebri

  • Seo, Jong-Hun;Seo, Hong-Ju;Kim, Seok-Won;Shin, Ho
    • Journal of Korean Neurosurgical Society
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    • v.49 no.5
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    • pp.296-298
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    • 2011
  • Spinal intradural cysticercosis is a rare manifestation of neurocysticercosis. We report a unique patient who showed visual symptoms and normal imaging of the brain caused by isolated spinal neurocysticercosis. A 59-year-old male patient was admitted to the emergency unit with a history of severe headache and progressive blurred vision. Brain computed tomographic scanning and magnetic resonance imaging showed normal cerebral anatomy without hydrocephalus. The fundoscopic evaluation by an ophthalmologist showed bilateral papilledema. Perimetry studies revealed visual field defects in both eyes. With the diagnosis of pseudotumor cerebri, a lumbar tap was attempted; however, we could not drain the cerebrospinal fluid in spite of appropriate attempts. Lumbar magnetic resonance imaging revealed multilevel intraspinal lesions that were confirmed histologically to be neurocysticercosis. An intraoperative lumbar puncture revealed an increased opening pressure and cytochemical analysis showed elevated cerebrospinal fluid protein level. The headache resolved immediately after surgery. However, the visual symptoms remained and recovered only marginally despite antihelminthic medications after six months of operation.

Relationship between the nucleolar cycle and chromatoid body formation in the spermatogenesis of $Phrynops$ $geoffroanus$ (Reptilia Testudines)

  • Peruquetti, Rita L.;Taboga, Sebastiao R.;Cabral, Silvia R.;De Oliveira, Classius;Azeredo-Oliveira, Maria T.
    • Animal cells and systems
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    • v.16 no.2
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    • pp.104-113
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    • 2012
  • The nucleolus is a distinct nuclear territory involved in the compartmentalization of nuclear functions. There is some evidence of a relationship between nuclear fragmentation during spermatogenesis and chromatoid body (CB) formation. The CB is a typical cytoplasmic organelle of haploid germ cells, and is involved in RNA and protein accumulation for later germ-cell differentiation. The goal of this study was to qualitatively and quantitatively describe the nucleolar cycle during the spermatogenesis of $Phrynops$ $geoffroanus$ (Reptilia Testudines), and compare this nucleolar fragmentation with CB formation in this species through the use of cytochemical and ultrastructural analysis. Qualitative analysis showed a fragmentation of the nuclear material after pachytene of the first meiotic division in the primary spermatocytes. Quantitative analysis of the nucleolar cycle revealed a significant difference in the number of nucleoli and in the size of the nucleolus between spermatogonia and early spermatids. Using ultrastructural analysis, we recorded the beginning of the CB formation process in the cytoplasm of primary spermatocytes at the same time as when nuclear fragmentation occurs. In the cytoplasm of primary spermatocytes, the CB was observed in association with mitochondrial aggregates and the Golgi complex. In the cytoplasm of early spermatids, the CB was observed in association with lipid droplets. In conclusion, our data show that the nucleolus plays a role in the CB formation process. During spermatogenesis of $P.$ $geoffroanus$, the CB is involved in some important biological processes, including acrosome formation and mitochondrial migration to the spermatozoon tail and middle piece region.

Distribution of Glutamate Receptors in the Retina of the Greater Horseshoe Bat (Rhinolophus ferrumequinum) (한국관박쥐 망막에서 글루타메이트 수용체의 분포 양상)

  • Kwon, Oh-Ju;Jeon, Chang-Jin
    • Journal of Korean Ophthalmic Optics Society
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    • v.19 no.3
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    • pp.413-418
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    • 2014
  • Purpose: The objective of this study was analyzing the distribution of the excitatory neurotransmitter glutamate receptor to investigate the function in the retina of the greater horseshoe bat. Methods: After retinal tissues of adult greater horseshoe bat were cut into $40{\mu}m$ vertical sections, standard immuno-cytochemical techniques was applied for analysis. Immunofluorescence images were obtained using the Bio-Rad MRC 1024 laser scanning confocal microscope. Results: AMPA (GluR1-4), Kainate (GluR5-7, KA1-2) and NMDA (1, 2A, 2B) mainly distributed in the inner plexiform layer and outer plexiform layer. KA1 receptors have existed not only plexiform layer but also ganglion cell layer. Conclusions: The greater horseshoe bat has same neuron and neurotransmitter to mammalian retina. These findings suggest that bat has a functional retina for visual analysis.

Ultrastructure of the Cell Wall of a Null Pigmentation Mutant, npgA1, in Aspergillus nidulans

  • Chung, Yun-Shin;Kim, Jung-Mi;Han, Dong-Min;Chae, Keon-Sang;Jahng, Kwang-Yeop
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.224-231
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    • 2003
  • The null pigmentation mutant (npgA1) of Aspergillus nidulans was previously characterized by its production of no pigment at any stage of its life cycle, its reduction in hyphal branching, and its delay in the asexual spore development. The chemical composition of the cell wall was also altered in npgA1 mutants that became more sensitive to Novozyme 234$\^$TM/, which is possibly due to a structural defect in the cell wall. To investigate the effects of the cell wall structure on these pleiomorphic phenomena, we examined the ultrastructure of the cell wall in the npgA1 mutant (WX17). Scanning electron micrographs (SEM) showed that after being cultured for six days, the outermost layer of the conidial wall of WX17 peeled off. Although this phenotype suggested that the cell wall structure in WX17 may be modified, examination using TEM of the fine structure of cross-sectioned hyphal wall of WX17 did not show any differences from that of FGSC4. However, staining for carbohydrates of wall layers showed that the electron-translucent layer of the cell wall was missing in WX17. In addition, the outermost layer H1 of the hyphal wall was also absent in WX17. The ultrastructural observation and cytochemical analysis of cell walls suggested that the pigmentation defect in WX17 may be attributed to the lack of a layer in the cell wall.

The Effect of Conjugated Linoleic Acid Isomers on the Cell Proliferation, Apotosis and Expressions of Uncoupling Protein (Ucp) Genes during Differentiation of 3T3-L1 Preadipocytes (Conjugated Linoleic Acid 이성체가 3T3-L1 지방전구세포 분화중 세포증식, 세포사멸 및 Ucp 유전자 발현에 미치는 영향)

  • Kwon So-Young;Kang Keum-Jee
    • Journal of Nutrition and Health
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    • v.37 no.7
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    • pp.533-539
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    • 2004
  • It has been reported that CLA decreases fat deposition in vivo and in vitro experiments. Among CLA isomers, c9t11 and t10c12 have been shown to exert active biological activities. For example, t10c12 reduces body weight and increases lean body mass, whereas, c9t11 has little effect on body fattness. However, the underlying molecular mechanism for the anti-obesity action of CLA isomers are not well understood. The purpose of this study was to examine the effects of t10c12 and c9t11 on lipid accumulation, cell proliferation, cell death and the expression levels of Ucp genes which are proposed as targets for anti-obesity in 3T3-L1 preadipocytes. Isomers of CLA at 50$\mu$M were added into preadipocyte differentiation medium for 3, 6 and 9days. Control cells received only the vehicle in the differentiation medium. Cytochemical analyses for lipid accumulation, cell proliferation and apotosis were carried out to compare lipidogenesis and cellular activity. RT-PCR analysis of GAPDH, Ucp 2,3 and 4 were also performed to find any modulatory effects of CLA isomers on the metabolic genes. Lipid accumulation indicated by Oil Red-O staining was inhibited in CLA isomers as compared to the control. T10c12 isomer showed less lipidogenesis than c9t11 did. A decrease occurred in CLA isomers as shown by BrdU incorporation. Apotosis has occured at higher level in t10c12 when compared to that of t9c11. Ucp 2, 3 and 4 genes were also upregulated in CLA isomers. T10c12 showed higher level of Ucp gene expressions than the c9t11 did. The biological activities of CLA isomers were also found to be different during differentiation of 3T3-L1 preadipocytes, suggesting that different isomers may be active in certain stage of lipidogenesis. The results indicate that both c9t11 and t10c12 CLA isomers decrease lipidogenesis, inhibit cell proliferation, increase cell death and upregulate in Ucp gene expressions during 3T3-L1 preadipocyte differentiation. T10c12 isomer was more effective than c9t11 in overall anti-obesity activity.

Effect of Exercise Training on Aging Atrophy in Rat Skeletal Muscle II. Effect of Long Term Weight-Training (흰쥐 골격근의 노화성 위축에 대한 운동훈련의 영향 - II. 장기간에 걸친 체중부하 훈련의 영향 -)

  • Park, Sung-Han;Park, Won-Hark;Lee, Yong-Deok;Kim, Jung-Ki
    • Applied Microscopy
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    • v.25 no.4
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    • pp.26-51
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    • 1995
  • The present study was designed to examine effect of long term weight-training on aging atrophy in the rat skeletal muscle. Male rats of 8, 15, and 24 month old were used. Each age groups included control and weight-training for 5 months by using body press apparatus. The histo- and cytochemical, ultrastructural and stereological changes in aging skeletal muscles of the rat were observed in the present study. During the training period the body weight and muscular weight in all groups except the rectus femoris and the gastrocnemius in young age groups remained constant, but muscular weights were increased in the rectus femoris and the gastrocnemius muscles in young age groups. In trained rat, the volume density of muscle fiber type IIA and IIB were increased, but those of type IIC was decreased. Type I remained constant in 8 and 15 month old age groups, but reduced in the tibialis anterior and the gastrocnemius muscles in the 24 month old groups. Some histotological and ultrastructural changes associated with age were found: numerical increase of cytiplasmic vacuoles, lysosomes, lipofuscins, and irregularity of myofibrils. At 24 month old groups some unusual formation of contraction band and muscle splitting were observed. After weight-training, ultrastructural degenerative changes occured in the type I muscle fiber, such as splitting of muscle fiber, disorganization of myofilaments, swelling of mitochondria, accumulation of many lipid droplets, appearance of many lysosomes and residual bodies and necrotic fibers, in the old age groups. But, in the type II muscle fibers hypertrophy of muscle fiber appeared without any noticible damage as the type I. The activities of $Mg^{++}$ -ATPase decreased with age and this enzyme activities in the trained rat were significantly decreased with age. Activities of the acid phosphatase were increased with age and significantly in the trained rat. In stereological analysis, volume density of the myofibrils and the tubular system were increased, on the other hand there mitochondrial capacity was decreased. These experimental results suggested that old rats are not susceptible to be affected by weight-training as young rats, and that physical capacity of the rats must be considered when old rats are exercised for training.

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